Authors used a form of rl lacking the C-terminal 52 amino acids that should be unable to bind substrates.
Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).
Two-hybrid system: yeast LexA-BD/B42-AD
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.
Interaction in vitro; proteins produced as a recombinant fusion protein in bacterial system.
Interaction in vitro; proteins produced as a recombinant fusion protein in bacterial system.
Purified proteins were photocross-linked to stabilize the complex, protease digested, then cross-linked peptides identified by mass spectrometry.
T198A, coordinates relative to rl-PC
Y200F, coordinates relative to rl-PC
Source was cell extracts of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.
Source was embryos of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.
